Antibodies, Proteins, Kits and Reagents for Life Science

368 544 ₸ Product size

100 µl 368 544 ₸

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Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)
Suitable for: ELISA, Dot blot, ICC/IF, WB, IHC-P, IP
Reacts with: Mouse, Human

Product name

Anti-Histone H3 (phospho S10 + T11) antibody [E173]
See all Histone H3 primary antibodies
Description

Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)
Host species

Rabbit
Specificity

This antibody detects Histone H3 phosphorylated on both Serine 10 and Threonine 11. However, the antibody shows higher affinity for phosphorylated Serine 10 than for phosphorylated Threonine 11. This was validated by ELISA, Dot Blot and WB peptide blocking experiments.

Tested Applications & Species

Application	Species
Dot	Recombinant fragment
ELISA	Recombinant fragment
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (phospho S10).
Database link: Q16695
Positive control
- WB: HeLa whole cell lysate (ab150035) treated with Calyculin A. IHC-P: Lymphoma tissue. ICC/IF: HeLa cells IP: HeLa treated with calyculin A whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Clonality

Monoclonal
Clone number

E173
Isotype

IgG
Research areas

Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/10000 dilution

Lane 1 : Untreated HeLa cell lysate
Lane 2 : HeLa cell lysate treated with calyculin A
Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 15 kDa
Observed band size: 17 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking and dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor^® 594). DAPI (blue) was used as a nuclear counterstain.
Confocal image showing nuclear staining on mitotic HeLa cell.
Immunoprecipitation - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Purified ab32107 at 1/50 dilution (2µg) immunoprecipitating Histone H3 in HeLa treated with calyculin A whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with calyculin A whole cell lysate 10µg
Lane 2 (+): ab32107 + HeLa treated with calyculin A whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32107 in HeLa treated with calyculin A whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 17 kDa
Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/1000 dilution

Lane 1 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho S10 peptide
Lane 2 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho T11 peptide
Lane 3 : HeLa whole cell lysate -treated with Calyculin A with unmodified Histone H3 peptide
Lane 4 : HeLa whole cell lysate -treated with Calyculin A with no peptide

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 1 second

Blocking/Diluting buffer 5% NFDM/TBST
Dot Blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Dot blot analysis of Histone H3 phospho S10 peptide (Lane 1), Histone H3 phospho T11 peptide (Lane 2) and Histone H3 unmodified peptide (Lane 3) labelling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) was used as the secondary antibody at a dilution of 1/10 000.

Blocking and diluting buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Ab32107, at a 1/50 dilution, staining Histone H3 (phospho S10 + T11) in paraffin embedded lymphoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) This image is courtesy of an Abreview submitted by Dr Kirk McManus

ab32107 (1/1000) staining Histone H3 (phospho S10 + T11) in paraformaldehyde-fixed, DAPI counterstained HeLa cells. Secondary antibody: Goat anti-Rabbit IgG conjugated to Cy3® (1/200). Please refer to abreview for further details.
See Abreview
ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (100 ng/mL).

Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).
ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (10 ng/mL).

Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).
Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

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